Abstract
Assays of plasminogen first require its conversion to the enzyme plasmin by common activators. The enzyme is then assayed on the basis of its esterolytic activity toward N-a-toluenesulfonyl-L-arginine methylester (TosArgOMe, TAME) utilizing a recording pH stat to titrate the amount of acid, Na-tosyl-L-arginine, liberated with time. In this chapter the Colorimetric method is described. This method is used if the equipment to perform esterolytic activity is not available. Colorimetric method is very sensitive and is used for analysis of the concentration of TosArgOMe remaining after incubation of plasmin with this substrate for a given period of time. This method is based upon the quantitative reaction of esters, such as TosArgOMe, with hydroxylamine, yielding the hydroxamic acid derivative. Upon addition of ferric chloride, a ferric ion-hydroxamic acid complex is formed, possessing spectral properties different from ferric chloride. The chapter also explains the Potentiometric Method. The purification procedure of plasminogen, chemical, physical and kinetic properties of rabbit plasminogen are also discussed.
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